中国动脉硬化杂志2018，Vol.26Issue(2)：115-121,7.
脂肪分化相关蛋白通过抑制中性胆固醇酯水解酶表达促进RAW264.7细胞内脂质积蓄
Adipophilin enhances intracellular lipid accumulation by inhibiting neutral cholesterol ester hydrolase in RAW264.7 macrophages
摘要
Abstract
Aim To explore the mechanism of adipophilin regulating neutral cholesterol ester hydrolase (nCEH) expression thus mediating lipid accumulation in RAW264.7 macrophages.Methods The constructed retroviral plasmid vectors with adipophilin of no expression and over expression were transfected into package cell PA317,then collected virus solution was used to infect RAW264.7 macrophages by screening and identifying to obtain RAW264.7 cell lines with adipophilin stably silenced and highly expressed.The mRNA and protein expression levels of nCEH were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively.High performance liquid chromatography (HPLC) was taken to detect the contents of intracellular cholesterol ester.Results The cholesterol and cholesterol ester levels were significantly higher in adipophilin over-expression group than in the control group (P＜0.01),while dramatically lower than that in the control group when adipophilin was supressed (P＜0.01).In addition,adipophilin inhibited nCEH expression.Furthermore,in RAW264.7 macrophages with high adipophilin expression,treatment with 100 nmol/L protein kinase Cδ agonist PMA for 30 min resulted in remarkable reductions of nCEH mRNA and protein expression(P＜0.05),on the contrary,treatment with 100 nmol/L PKCδ inhibitor Rottlerin for 30 min increased the expression of nCEH(P＜0.05).Conclusion Adipophilin may exert influence over lipid accumulation by inhibiting the expression of nCEH in RAW264.7 macrophages,and PKCδ played an important role in this regulatory mechanism.关键词
脂肪分化相关蛋白/中性胆固醇酯水解酶/蛋白激酶Cδ/动脉粥样硬化Key words
Adipophilin/Neutral cholesterol ester hydrolase/Protein kinase Cδ/Atherosclerosis分类
医药卫生引用本文复制引用
袁旭,孟磊,陈雁斌,张瑞,袁中华..脂肪分化相关蛋白通过抑制中性胆固醇酯水解酶表达促进RAW264.7细胞内脂质积蓄[J].中国动脉硬化杂志,2018,26(2):115-121,7.基金项目
国家自然科学基金资助项目(30971268) （30971268）
